The Derived Components of Gnaphalium hypoleucum DC. Reduce Quorum Sensing of Chromobacterium violaceum.

Gnaphalium hypoleucum DC. was first recorded in the Chinese National Pharmacopoeia "Yi Plant Medicine". There is no detailed report on its main components' activity in suppressing the quorum sensing activity (QS) of bacteria. Our study aimed to screen the main components in extracts of G. hypoleucum DC. in order to measure their effects on bacterial QS activity and to explore specific quorum sensing mechanisms that are affected by G. hypoleucum DC. extracts. Crude extracts of G. hypoleucum DC. contained significant amounts of two compounds shown to inhibit bacterial QS activity, namely apigenin and luteolin. Apigenin and luteolin in crude extracts of G. hypoleucum DC. showed substantial inhibition of pigment formation, biofilm production, and motility in Chromobacterium violaceum ATCC 12472 compared to the effects of other phytochemicals from G. hypoleucum DC. Apigenin and luteolin exhibited a strong QS inhibitory effect on C. violaceum, interfering with the violacein pigment biosynthesis by downregulating the vioB, vioC, and vioD genes. In the presence of signal molecules, the QS effect is prevented, and the selected compounds can still inhibit the production of the characteristic purple pigment in C. violaceum. Based on qualitative and quantitative research using genomics and bioinformatics, we concluded that apigenin and luteolin in crude extracts of G. hypoleucum DC can interfere with the generation of QS in C. violaceum by downregulating the vioB, vioC, and vioD genes. Indeed, G. hypoleucum DC. is used for the treatment of bacterial infections, and this research provides new ideas and potential alternative uses for medicinal plants.

Streptomyces blattellae, a novel actinomycete isolated from the in vivo of a Blattella germanica.

During a screening for novel and useful actinobacteria in desert animal, a new actinomycete was isolated and designated strain TRM63209T. The strain was isolated from in vivo of a Blattella germanica in Tarim University in Alar City, Xinjiang, north-west China. The strain was found to exhibit an inhibitory effect on biofilm formation by Candida albicans ATCC 18,804. The strain was observed to form abundant aerial mycelium, occasionally twisted and which differentiated into spiral spore chains. Spores of TRM63209T were observed to be oval-shaped, with a smooth surface. Strain TRM63209T was found to grow optimally at 28 °C, pH 8 and in the presence of 1% (w/v) NaCl. The whole-cell sugars of strain TRM63209T were rhamnose ribose, xylose, mannose, galactose and glucose, and the principal polarlipids were found to be diphosphatidylglycerol, phos-phatidylethanolamine, phosphatidylcholine, phosphatidylinositol mannoside, phosphatidylinositol and an unknown phospholipid(L). The diagnostic cell wall amino acid was identified as LL-diaminopimelic acid. The predominant menaquinone was found to be MK-9(H6) (14.64%), MK-9(H2) (19.65%), MK-9(H8) (22.34%), MK-10(H2) (25.37%). The major cellular fatty acids were identified as iso-C16:0, 16:0, anteiso-C15:0, anteiso-C17:0, iso-C15:0 and Sum in Feature 3. Analysis of the 16S rRNA sequence showed that strain TRM63209T exhibits high sequence similarity to Streptomyces bungoensis strain DSM 41781T 98.20%. A multi-locus sequence analysis of five house-keeping genes (atpD, gyrB, rpoB, recA and trpB) and phylogenomic analysis also illustrated that strain TRM63209T should be assigned to the genus Streptomyces. The DNA G + C content of the strain was determined to be 70.2 mol%. Average nucleotide identity (ANI) between strain TRM63209T and S. bungoensis DSM 41781T, Streptomyces phyllanthi PA1-07T, Streptomyces longwoodensis DSM 41677T and Streptomyces caeruleatus NRRL B-24802T were 82.76%, 82.54%, 82.65%, 84.02%, respectively. Digtal DNA-DNA (dDDH) hybridization were 26.30%, 25.10%, 26.20%, 29.50%, respectively. Therefore, it is concluded that strain TRM63209T represents a novel species of the genus Streptomyces, for which the name Streptomyces blattelae is proposed. The type strain is TRM63209T (CCTCC AA 2018093T = LMG 31,403 = TRM63209T).

[Medication experience of TCM Master Academician Wang Qi in the treatment of premature ejaculation: Data mining and analysis].

Objective: To analyze the academic thought, medication experience and prescription rules of Academician Wang Qi in the treatment of premature ejaculation (PE) using the TCM inheritance support platform (V2.5). METHODS: We collected and sorted out the medical records on the treatment of PE from Academician Wang Qi's Clinic. We established a database of medical records on the TCM inheritance support platform, analyzed the drugs and prescriptions in the database and explored new prescriptions using "statistical reports" and "data analysis" systems on the platform. RESULTS: A total of 91 effective prescriptions were recorded, involving 148 TCM drugs, with Phellodendron, Amomum Villosum, Polygala Tenuifolia, Tuckahoe, Lodestone, Oyster, Acanthopanax Senticosus, Uncaria, Tribulus, and Keel as the top 10 with the highest frequency of use, which were featured mainly by "warm" and "cold" concerning the four natures, "sweet", "bitter" and "pungent" relating to the five flavors, and acting on "kidney meridian", "liver meridian" and "heart meridian" in terms of the meridian tropisms. In addition, 5 new prescriptions were obtained through unsupervised entropy hierarchical clustering. CONCLUSIONS: In the treatment of PE, Academician Wang Qi employs tranquilizing the mind and consolidating the kidney (An Zhi Gu Shen) as the primary strategy, taking into account the three organs of heart, liver and kidneys, focusing on the phase of calming the mind or regulating the liver or clearing the kidney or controlling fire, and adding or reducing drugs according to different conditions and syndromes, which conforms to his diagnosis and treatment mode of "body differentiation－disease differentiation－syndrome differentiation". The analysis of the potential new prescriptions also accords with Academician Wang Qi's rules of medication, which can provide some ideas for the clinical treatment of and scientific researches on premature ejaculation in the future.

[Academician Wang Qi's medication rules for oligoasthenozoospermia: An analysis based on the Traditional Chinese Medicine Inheritance Auxiliary Platform].

OBJECTIVE: To analyze the medication rules for oligoasthenozoospermia (OAZ) observed by Wang Qi, an academician, master of Traditional Chinese Medicine (TCM) and initiator of andrology in TCM. METHODS: We collected the outpatient cases of OAZ treated by Wang Qi and established a database of clinical medical records using the TCM Inheritance Auxiliary Platform. Employing the integrated rule-based system for analysis of the software, we modified the mutual information method, complex system entropy clustering analysis and other data mining methods, and summarized the medication rules Wang Qi followed in the treatment of OAZ. RESULTS: A total of 134 prescriptions made by Wang Qi for the treatment of OAZ were collected, involving 110 TCM drugs, which are mainly neutral and warm in nature and taste sweet and mostly act through the liver and kidney meridians. The core formula ingredients of the prescriptions included Morinda officinalis, Cuscuta chinensis, Lycium barbarum, Mulberry, Angelica sinensis, Astragalus mongholicus and Fish Maw, and most frequently Morinda officinalis, Cuscuta chinensis, Lycium barbarum and Mulberry. CONCLUSIONS: Wang Qi holds that kidney deficiency, dampness-heat, blood stasis and toxin are the main pathogenic factors for OAZ. The basic treatment of OAZ is to invigorate the kidney and replenish the essence, and meanwhile activate blood circulation, dissipate stasis and eliminate dampness-heat.

[Bioavailability of Dissolved Organic Carbon in Rivers for Typical Vegetation Types in the Permafrost Regions on the Qinghai-Tibet Plateau].

Samples collected from 12 rivers with typical vegetation types in the permafrost regions on the Qinghai-Tibetan Plateau were incubated in the laboratory, and the relationships among the vegetation types, river discharges, the compositions of dissolved organic carbon (DOC), permafrost areas, riverine DOC concentration, biodegradability of dissolved organic carbon (BDOC), and the biodegradation kinetics were examined. The results showed that the DOC concentrations of typical vegetation types in the basin, such as alpine meadow (AM), alpine swamp meadow-alpine meadow (ASM-AM), alpine meadow-alpine steppe (AM-AS), and alpine meadow-alpine steppe-bare soil (AM-AS-BL), were (5.17±0.21), (5.02±0.50), (3.55±0.25), and (2.79±0.41) mg ·L-1, respectively. The values for the bioavailability of river DOC of different vegetation types were (23.54±2.62)%, (23.66±3.31)%, (18.17±5.26)%, and (11.72±15.56)%, respectively. Correspondingly, the riverine DOC aromaticity increased along with the vegetation cover, while the biodegradation and degradation rates decreased gradually. During the incubation, the reaction of BDOC was in accordance with the first-order kinetics equation. Furthermore, the BDOC in continuous permafrost regions of the rivers was greater than that in the non-continuous permafrost regions. The BDOC in higher discharges were lower than those with lower discharges. Taken together, the results suggested that the vegetation types were the main controlling factors for the BDOC, and BDOC was also related to the discharge and permafrost.

[Inhibitors of JAK2/STAT Signaling Pathway and Hematologic Malignancies-Review].

Jauns kinase (JAK)/transducer and activator of transcription（STAT） pathway is a classical approach to study the rapid changes of the gene expression in specific target cells by a variety of extracellular signals. The JAK and STAT transfer cytokine receptor signaling plays a unique role in multiple cellular and molecular biological changes.The abnormal signal of JAK/STAT pathway will lead to the hematopoietic abnormalities.Studies had shown that the abnormal activation of JAK2/STAT signaling pathway are in many kinds of malignant hematological diseases, such as in acute lymphoblastic/myeloid leukemia, chronic myeloid leukemia, lymphoma, myelodysplastic syndromes, myeloprofilerative neoplasm, especially in the patients of myeloproliferative neoplasm(MPN) with JAK gene mutation(JAK2V617F), this mutation has an important value for MPN diagnosis. At present, the effect of the specific inhibitors of JAK2 has showed good perspective, which had been applied to clinic treatment and achieved remarkable curative effect. In this review, the JAK2/STAT signaling transduction, the JAK2 signal and hematologic malignancies, the kagulation of signaling pathway and the inhibitors of JAK2/STAT signaling pathway are summarized.

[Effect of IFN-α2b on COX-2 and Angiogenesis in JAK2V617F Mutation Myeloproliferative Neoplasms].

OBJECTIVES: To investigate the influence of interferon-alpha-2b (IFN-α2b) with JAK2 kinase, COX-2 and microvessel density in patients of MPN and the relation of JAK2V617F and COX-2 in human erythroleukemia cell line (HEL) cells. METHODS: Forty-two cases of MPN patients with JAK2V617F mutation of initial treatment were collected from the Frist hospital of Baoding, including the IFN-α2b treatment group with 17 cases and untreated group with 25 cases. 10 cases of idiopathic immune thrombocytopenic purpura (ITP) patients synchronization were enrolled as controls. JAK2V617F/JAK2 mutation burden of MPN patients was detected by real time PCR (qRT-PCR);the expression levels of p-JAK2, COX-2 and microvascular density (MVD) marked with CD105 inpathological tissues of bone marrow in patients of MPN and ITP were detected by immunohistochemistry. The HEL cells were treated with different concentrations of IFN-α2b. The cell proliferation inhibition rate was calculated by CCK-8 test;the apoptosis rate was detected by flow cytometry; cell migration ability was tested by transwell chambers. JAK2 and COX-2 mRNA were detected by semi-quantitative PCR; p-JAK2 and COX-2 protein in HEL cells were detected by Western blotting. RESULTS: The expression levels of p-JAK2, COX-2 protein and MVD in untreated group were significantly higher than those of control groups. p-JAK2, COX-2 and MVD levels were significantly reduced in patients treated with IFN-α2b. Cell growth inhibition rates and apoptosis rates raise up by dose of IFN-α2b in HEL cells at 48 h.The mRNA expression levels of JAK2 and COX-2 as well as protein expression levels of p-JAK2 and COX-2 had a decreasing tendency with the increase of IFN-α2b concentration at 48 h.The migration capacity level of HEL cells which treated with 0.5×10 4 U/L IFN-α2b after 24 h was lower than that of control group. CONCLUSIONS: Angiogenesis of MPN and COX-2 were inhibited by IFN-α2b which regulates JAK2 signal pathway.

[The Effect of Ruxolitinib on the Expression of VEGF and HIF-1α in Leukemia HEL Cells].

OBJECTIVES: To investigate the effect of Ruxolitinib on the expression of vascular endothelial growth factor (VEGF) and hypoxia inducible factor-1 α (HIF-1α) in HEL cells. METHODS: he HEL cells were treated with Ruxolitinib in different concentrations (1 nmol/L, 5 nmol/L, 10 nmol/L, 50 nmol/L, 100 nmol/L, 500 nmol/L). The growth inhibition of Ruxolitinib on HEL cells was detected by CCK-8 assay;the mRNA expression level ofJAK2 were measured by RT-PCR and the protein level of p-JAK2, VEGF, HIF-1α were observed by Western blot after treated with Ruxolitinib for 24,48,72 h. Chick chorioallantoic membrane (CAM) test was used to testify the effect of Ruxolitinib on angiogenesis. RESULTS: Ruxolitinib with different concentrations could inhibit HEL cells proliferation. RT-PCR showed that the mRNA level ofJAK2 decreased in a concentration-dependent manner and Western blot demonstrated that the expression levels of p-JAK2, VEGF and HIF-1α were lower in Ruxolitinib treatment groups than those in control group (P<0.05) after HEL cells were treated with different concentrations of Ruxolitinib for 24,48,72 h. Ruxolitinib significantly suppressed blood vessels'formation in CAM. CONCLUSIONS: Ruxolitinib can inhibit VEGF, HIF-1α expression and angiogenesis of HEL leukemia cells by inhibiting JAK2 pathway.